Continuous Microfluidic Fabrication of Synthetic Asymmetric Vesicles
Award Abstract # 1429448
MRI: Development of a Microfluidic Instrument for High-throughput Production of Asymmetric Vesicles to Support Membrane Biology Research
| NSF Org: | DBI Div Of Biological Infrastructure |
| Awardee: | |
| Initial Amendment Date: | August 26, 2014 |
| Latest Amendment Date: | August 26, 2014 |
| Award Number: | 1429448 |
| Award Instrument: | Standard Grant |
| Program Manager: | Robert Fleischmann rfleisch@nsf.gov (703)292-7191 DBI Div Of Biological Infrastructure BIO Direct For Biological Sciences |
| Start Date: | September 1, 2014 |
| End Date: | August 31, 2018 (Estimated) |
| Total Intended Award Amount: | $294,000.00 |
| Total Awarded Amount to Date: | $294,000.00 |
| Funds Obligated to Date: | |
| History of Investigator: |
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| Awardee Sponsored Research Office: | 4400 VESTAL PKWY E BINGHAMTON NY US 13902-6000 (607)777-6136 |
| Sponsor Congressional District: | |
| Primary Place of Performance: | NY US 13902-6000 |
| Primary Place of Performance Congressional District: | |
| Unique Entity Identifier (UEI): | |
| Parent UEI: | |
| NSF Program(s): | Major Research Instrumentation |
| Primary Program Source: | |
| Program Reference Code(s): | |
| Program Element Code(s): | |
| Award Agency Code: | 4900 |
| Fund Agency Code: | 4900 |
| Assistance Listing Number(s): | 47.074 |
ABSTRACT The instrument will produce physiologically relevant synthetic vesicles to enable fundamental membrane biology research. The capabilities of the instrument will be demonstrated by using the synthetic vesicles to study the role of outer membrane vesicles in biofilm formation. Bacterial biofilms are ubiquitous in nature and have a large impact on human health and industry. It is imperative to understand the processes that contribute to the development of biofilm structure so that they can be exploited to control biofilm growth. This objective would not be achievable using traditional liposomes because they do not adequately mimic natural vesicles. Therefore they cannot be used to study complex phenomena where subtle differences in lipid composition and architecture are important. Emerging technologies are unable to synthesize uniform, unilamellar, asymmetric vesicles with controlled size at high throughput. The ability of the instrument to produce synthetic vesicles possessing all of these features is a paradigm shifting advancement in membrane biology and biofilm research. Long term, the vesicles built using the instrument have the potential to be used in vaccine development and as delivery vehicles for antimicrobial agents. As part of the instrument development, important fundamental issues relevant to microscale multiphase fluid flows will also be addressed. These include: flow focusing and interfacial stability, timescales for lipid self assembly, and emulsion kinematics.
An award is made to the State University of New York at Binghamton to develop a high throughput microfluidic instrument for constructing customizable vesicles with asymmetric lipid distributions. Such vesicles are superior to existing liposomes because they can be tailored to exactly replicate natural membranes and are therefore more physiologically relevant. The innovations provided by the instrument will have a broad impact on fundamental biofilm and membrane biology research. In particular, the physiologically relevant vesicles will enable studies that are impossible with current liposome technology. The interdisciplinary nature of this project provides an excellent training opportunity for students at all levels. Biologists and engineers often do not possess even the most fundamental skills in the others' discipline, limiting the ability to take on important research questions. This gap will be addressed by providing students with the opportunity to develop their interdisciplinary skills while contributing to the objectives of this project. Students at all levels will integrate with their cross disciplinary colleagues in the planning and execution of experiments and receive training in the laboratory techniques used by their research counterparts. To support interdisciplinary skills development in the wider community, a website will be created that provides instruction on protocols relevant to microfluidics, biofilms, and general laboratory practice. This website will be targeted to individuals with no background in the relevant discipline. It will also be used to disseminate the new instrument. This project will assist in establishing the formation of a Biofilm Microfluidics Initiative at SUNY Binghamton, which will tackle fundamental questions in the biofilms community with the aid of novel microfluidic tools.
PUBLICATIONS PRODUCED AS A RESULT OF THIS RESEARCH
Note: When clicking on a Digital Object Identifier (DOI) number, you will be taken to an external site maintained by the publisher. Some full text articles may not yet be available without a charge during the embargo (administrative interval). Some links on this page may take you to non-federal websites. Their policies may differ from this site.
L. Lu, R. Irwin, M. Coloma, J. Schertzer, and P. Chiarot "Removal of Excess Interfacial Material from Surface-modified Emulsions Using a Microfluidic Device with Triangular Post Geometry" Microfluidics and Nanofluidics , v.18 , 2015 , p.1233 10.1007/s10404-014-1521-9
L. Lu, J. Schertzer, and P. Chiarot "Continuous Microfluidic Fabrication of Synthetic Asymmetric Vesicles" Lab on a Chip , v.15 , 2015 , p.3591 10.1039/C5LC00520E
L. Lu, W. Doak, J. Schertzer, and P. Chiarot "Membrane Mechanical Properties of Synthetic Asymmetric Phospholipid Vesicles" Soft Matter , 2016 10.1039/C6SM01349J
PROJECT OUTCOMES REPORT
Disclaimer
This Project Outcomes Report for the General Public is displayed verbatim as submitted by the Principal Investigator (PI) for this award. Any opinions, findings, and conclusions or recommendations expressed in this Report are those of the PI and do not necessarily reflect the views of the National Science Foundation; NSF has not approved or endorsed its content.
A high-throughput microfluidic instrument for constructing custom synthetic vesicles with asymmetric membranes was developed and built as part of this project. This instrument has the potential to transform membrane biology research because all natural membranes are asymmetric (i.e. each leaflet of the bilayer contains a different lipid composition). The instrument provides simultaneous control over every aspect of vesicle quality: membrane lipid asymmetry, luminal content, unilamellarity, size over several orders of magnitude, uniformity, and throughput. Membranes with numerous architectures were created, including asymmetric phospholipid-phospholipid and phospholipid-lipopolysaccharide bilayers. The instrument provides membrane asymmetries as high as 95% immediately after vesicle formation. Over 80% of the vesicles remain stable for at least 6 weeks and the asymmetry is maintained for over 30 hours. The vesicles can be reliably transferred from the instrument and used for fundamental biofilm research, vaccine development, or other novel applications.
Synthetic vesicles built with this instrument were used to investigate the properties of bilayer membranes with varying lipid architectures. The mechanical properties of a membrane significantly affects its biological behavior. However, few studies had previously considered how trans-bilayer lipid asymmetry influenced membrane mechanical properties. Using our novel strategy for building asymmetric synthetic vesicles, our team discovered new effects on how lipid bilayer architecture governs the mechanical properties of biological membranes. Most notably, we were among the first to report on the impact of trans-bilayer asymmetry on the area expansion modulus of synthetic membranes. This was achieved using a customized micropipette aspiration system. We found that the bending modulus and area expansion modulus of synthetic asymmetric bilayers were up to 50% larger than the values acquired for symmetric bilayers. This was caused by the dissimilar lipid distribution in each leaflet of the bilayer for the asymmetric membrane. Since the mechanical properties of bilayer membranes play an important role in numerous cellular processes, these findings have significant implications for membrane biology and synthetic biology studies.
Graduate students in mechanical engineering and biology played an important role in the development of the microfluidic instrument. By participating in this project, these students had the opportunity to develop their interdisciplinary skills while contributing to the major goals. Videos were created that demonstrated the key protocols associated with the construction and operation of the instrument and are available to the public.
Last Modified: 11/29/2018
Modified by: Paul Chiarot
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